Quantitative determination of four immunosuppressants by high resolution mass spectrometry (HRMS)
Quantitative determination of four immunosuppressants by high resolution mass spectrometry (HRMS)
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高分辨质谱法（HRMS）对四种免疫抑制剂的定量测定
DOI：
10.1515/cclm-2015-0863
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发表时间：
2016-07-01
影响因子：
6.8
通讯作者：
Lackner, Karl J.
中科院分区：
文献类型：
--
作者：
Bruns, Kai;Moennikes, Rene;Lackner, Karl J.
背景：利用三重四极杆仪器的液相色谱-串联质谱法（LC-MS/MS）已广泛应用于临床实验室内源性化合物、药物或代谢物的定量分析。相比之下，高分辨率质谱（HRMS）由于其有限的动态范围而通常用于化合物鉴定。最近，具有增强线性动态范围的HRMS仪器已经可用。本研究的目的是评估HRMS在临床实验室的快速定量应用。方法：建立高通量UPLC-TOF-MS同时定量环孢素A、他克莫司、西罗莫司和依维莫司的方法。所有免疫抑制剂使用Agilent 6540 Q-TOF系统在tof模式下作为钠加合物进行分析。从全扫描数据中提取分析物的离子色谱图和内标。根据CLSI建议，对该分析进行评估并与已建立的LC-MS/MS分析进行比较。结果：新型HRMS法总运行时间为3 min。该检测在所有四种免疫抑制剂的临床相关浓度范围内呈线性。方法与建立的LC-MS/MS分析的相关性在R-2 = 0.99和R-2 = 0.97之间。三个相关浓度水平的总变异系数（CV T）分别为4.5% ~ 6.4%（他克莫司）、7.4% ~ 8.0%（-西罗莫司）、8.0% ~ 8.8%（依维莫司）和6.1% ~ 7.4%（环孢素A）。结论：高分辨率TOF-MS与LC-MS/MS对环孢素A、他克莫司、西罗莫司和依维莫司的监测具有相同的定量效果。HRMS有可能在临床实验室取代传统的LC-MS/MS，因为它简化了分析开发（不需要优化片段和产物离子），其全扫描数据可以提供额外的信息。
Background: Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) utilizing triple-quadrupole instruments has been widely used for quantification of endogenous compounds, drugs or metabolites in clinical laboratories. In contrast, high-resolution mass spectrometry (HRMS) is typically used for compound identification due to its limited dynamic range. Recently HRMS instruments with enhanced linear dynamic range have become available. The aim of this study was to evaluate HRMS for fast quantitative applications in a clinical laboratory.Methods: A high throughput UPLC-TOF-MS assay for simultaneous quantification of cyclosporin A, tacrolimus, sirolimus and everolimus was developed. All immunosuppressants were analyzed as sodium adducts in TOF-only mode using an Agilent 6540 Q-TOF system. Extracted ion chromatograms of analytes and internal standards were created from full-scan data. The assay was evaluated and compared to an established LC-MS/MS assay according to CLSI recommendations.Results: The novel HRMS assay has a total run time of 3 min. The assay is linear in a clinical relevant concentration range for all four immunosupressants. Method correlations vs. established LC-MS/MS assay were between R-2 = 0.99 and R-2 = 0.97. Total coefficients of variation (CV T) ranges were 4.5%-6.4% (tacrolimus), 7.4%-8.0% (-sirolimus), 8.0%-8.8% (everolimus) and 6.1%-7.4% (cyclosporine A) for three relevant concentration levels each.Conclusions: High resolution TOF-MS and LC-MS/MS show equivalent quantitative performance for monitoring of cyclosporin A, tacrolimus, sirolimus and everolimus. HRMS has the potential to replace conventional LC-MS/MS in clinical laboratories because it simplifies assay development (no optimization of fragmentations and product ions necessary) and its full-scan data can provide additional information.